Human papillomavirus (HPV), the most prevalent sexually transmitted infection and a leading cause of cervical cancer, often lacks rapid and cost-effective diagnostic methods. This study introduces a novel colorimetric biosensor for specific and sensitive HPV18 detection. This biosensor was developed using an in silico-designed Gquadruplex aptamer, chosen for its inherent stability and specific binding capabilities, in conjunction with gold nanoparticles (AuNPs). Our in silico design identified APT43 as the top candidate aptamer, demonstrating high binding affinity and specificity for the HPV18 L1 protein via molecular docking. AuNPs were then experimentally synthesized and functionalized with APT43. Successful conjugation was confirmed by DLS characterization, evidenced by a hydrodynamic size increase from similar to 29 nm to similar to 36 nm and a zeta potential shift from -36 mV to -29.8 mV. The developed biosensor exhibited high specificity for HPV18, showing no cross-reactivity with types 16, 11, or 6. It also achieved a remarkably low detection limit of 85 viral copies/mL, significantly lower than the typical similar to 1000 copies/mL reported for many commercial PCR kits. This rapid, visual, aptamer-induced AuNP aggregation system offers a sensitive, specific, and cost-effective diagnostic tool for HPV18, particularly beneficial in resource-limited settings. This work reports the first computationally designed aptamer capable of typespecific discrimination of HPV18 from other high- and low-risk HPV types in real clinical samples, marking a significant advancement in rapid HPV diagnostics.

Equine herpesvirus type 1 (EHV-1) is a globally prevalent equine pathogen responsible for severe respiratory, neurological, and reproductive disorders. Accurate and ultrasensitive detection of EHV-1 is critical for timely disease management. In this study, we report the development of the first G-quadruplex-forming aptamer specifically designed for EHV-1 detection. The aptamer was generated using an in silico approach, and its G-quadruplex conformation was confirmed using circular dichroism (CD) spectroscopy and crystal violet fluorescence assays. Binding affinity and specificity were assessed using a comprehensive panel of analytical techniques, including colorimetric assays, enzyme-linked apta-sorbent assay (ELASA), surface plasmon resonance (SPR), CD spectroscopy, and fluorescence analysis. The aptamer exhibited a high binding affinity in the picomolar range, as determined by SPR. In both colorimetric and ELASA platforms, it enabled the detection of as few as 10 viral particles per milliliter, compared to the 1000 viral particles per milliliter required by conventional PCR. ELASA results demonstrated excellent diagnostic performance, yielding an area under the curve of 0.96. Importantly, this aptamer-based method eliminates the need for DNA extraction, primers, or gel electrophoresis. These findings underscore the aptamer's strong potential as a cost-effective, rapid, and user-friendly point-of-care diagnostic tool for EHV-1, especially in low-resource or field settings.

Fire blight, caused by Erwinia amylovora, is a significant threat to fruit crops, with limited biocontrol methods. This study aimed to develop a nanosystem using mesoporous silica nanoparticles (MSNs) loaded with a phenolic plant extract (ZP) derived from Myrtus communis, Thymus vulgaris, and Curcuma longa, and coated with natural biopolymers Gum Tragacanth (GT) and sodium alginate (SA). The MSNs were synthesized and characterized by XRD, FTIR, and TEM, exhibiting a specific surface area of about 750 m(2)/g and an average pore diameter of 5 nm. ZP was effectively loaded into the MSNs with a loading efficiency of similar to 25 %, and GT-MSNs-ZP demonstrated sustained release, releasing 56 % of phenolic compounds over 168 h. In antibacterial tests, GT-MSNs-ZP demonstrated the highest effectiveness against E. amylovora, maintaining inhibition for up to 7 days. In vivo experiments showed that GT-MSNs-ZP reduced diseased leaves by 60 % at a concentration of 5/1000 mL/mL, comparable to commercial pesticides. Additionally, the system showed no adverse effects on beneficial bacteria such as Rhizobium meliloti and Bacillus licheniformis. These results emphasize the potential of GT-MSNs-ZP as a sustainable and effective biocontrol solution for agricultural applications.

Pseudomonas syringae is a gram-negative bacterium that causes a diversity of diseases in numerous plants. Strategies to inhibit P. syringae growth include protective procedures; however, controlling the disease is complicated due to its rapid spread. Several antimicrobial agents can prevent this disease, such as chemical compounds, biological agents, secondary metabolites, nanoparticles, bacteriophages, and antimicrobial peptides (AMPs). The most effective way to control the disease is through chemical control. Using copper compounds and antibiotics is a conventional practice to decrease canker disease symptoms. However, due to environmental pollution caused by chemicals and bactericides and the resistance of different pathovars of P. syringae, other methods for bacterial pathogens control are needed. Biological control, using antagonistic bacteria has shown promising results against P. syringae under in vitro conditions. New studies focus on using secondary metabolites from plants to control plant diseases. Studies have shown that essential oils when preserved from degradation and evaporation by nanoparticles like mesoporous silica, can increase their antibacterial activities. Using nanoparticles, especially silver, is a suitable strategy for controlling P. syringae. However, high concentrations of silver nanoparticles are toxic. Bacteriophages and AMPs are recommended as alternatives to control bacterial infections in agriculture, including P. syringae. Combined treatments of phages and secondary metabolites have shown higher efficacy, potentially overcoming resistance. However, bacteriophages and AMPs are expensive and limited. In the end, using secondary metabolites and nanoparticles at low concentrations presents economic benefits and antibacterial activities without phytotoxic properties.

This study aimed to develop a predictive model for mucus-binding proteins using machine learning and to experimentally evaluate the anti-cariogenic effects of selected probiotic strains. In silico, a computational method was established utilizing Support Vector Machine (SVM) and AdaBoost algorithms with pseudo amino acid composition (PseAAC) for protein sequence representation. The predictive model achieved high accuracy. Specifically, the SVM model demonstrated 94

Background Nitrate and acrylamide as carcinogenic substances are increased during the baking process of foods, such as cereals.Objective This study aimed to reduce the amount of acrylamide and nitrate in three types of cereals, wheat, barley, and maize, by treatment with probiotic bacteria and several plant extracts.Methods Three types of plant extracts were prepared from Coriandrum sativum, Nigella sativa, and Thymus sp. leaves and stem. Also, Lactobacillus casei subspecies rhamnosus LCR6013 was used as probiotic bacteria for bacterial treatment. Acrylamide and nitrate were measured by HPLC and UV-vis spectrophotometry.Results Adding plant extracts and LCR 6013 bacteria could reduce the level of nitrate and acrylamide in the cereal samples. Among plant extracts, nigella could reduce nitrate in all samples below detectable levels. Also, it was effective in reducing acrylamide content from samples to the extent of 87% in barley, 60% in wheat, and 100% in corn. Bacterial treatment could also reduce nitrate levels between 70 and 100% while having a variable impact on decreasing acrylamide. One-way analysis of variance (ANOVA) was used to determine statistically significant results.Conclusion It was concluded that pre-baking exposure to plant extract and bacteria is effective in the reduction of nitrate and acrylamide quantity in the heat processing of cereals.

Background: Therapeutic effects of plant metabolites have been used for the treatment of burns, wounds and infections over the centuries. Electrospun nanofibers containing plant metabolites have also been considered recently for the development of new and efficient wound dressings. Ferula assa-foetida has received much attention in traditional medicine due to its numerous healing properties. Objective: In the present study, polyvinyl alcohol (PVA) nanofibers containing aqueous extracts of F. assa-foetida gum (FAE) were prepared and characterized. The antibacterial activity of nanofibers was investigated. Methods: Electrospinning was utilized for the fabrication of PVA/FAE nanofibers. The morphology, physical and chemical properties of the synthesized nanofibers were investigated by scanning electron microscope (SEM), Fourier transform infrared (FTIR) spectroscopy, and contact angle test. Results: The uniform nanofibers with the average diameter of 256 nm were obtained by using 8 wt.% PVA, 1:4 (w: w %) ratio of PVA/FAE, needle to collector distance of 13 cm, 20 kV voltage, collector rotation speed of 3 m/min, and flow rate of 0.5 mL/h. The use of FAE led to the increased diameter of nanofibers and their contact angle compared to PVA nanofibers. Interestingly, the PVA/FAE nanofibers displayed considerable antibacterial activity against Escherichia coli and Staphylococcus aureus. Conclusion: The overall results indicated that PVA/FAE nanofibers can be considered as a potential candidate for the preparation of wound dressings with antibacterial properties.

Herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), which cause oral and genital herpes in humans, are prevalent worldwide. ELISA, real-time PCR, and cytological assays are conventional methods for detecting these viruses, but they are expensive and time-consuming. The main purpose of this study was to design a specific G-quadruplex aptamer for the simple and rapid detection of HSV-1 and HSV-2. In this study, a specific aptamer was designed using bioinformatics tools for binding to the glycoprotein gD in HSV-1 and HSV-2. After evaluating the binding of the aptamer to gD, based on the stability scores of the secondary and tertiary structures and molecular docking, the aptamer AptNR88 was selected, and its binding to the target protein was confirmed experimentally using a colorimetric system with gold nanoparticles. Gold nanoparticles with an average size of 30 nm were synthesized, and the AptNR88 was coated on them through hydrogen bonds and electrostatic interactions. The concentration of AptNR88 was subsequently optimized for resistance against salt-induced aggregation. The color changes of gold nanoparticles from red to purple due to salt aggregation were observed only in the presence of the AptNR88-virus complex after 15 min. These results confirmed the specific binding of AptNR88 to the gD of HSV-1 and HSV-2. The limit of detection (LOD) for AptNR88 was approximately 11.7 copies/ml for HSV-1 and 15.7 copies/ml for HSV-2. These results indicate that the designed aptamer for detecting these two viruses is sufficiently sensitive and specific.

Background: This study explores repetitive Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) sequences from the archaea Acidianus sp. and Acidianus ambivalens (A. ambivalens), as well as from the bacterium Yersinia ruckeri (Y. ruckeri). These sequences are compared with human microRNA (miRNA) sequences to investigate potential genetic similarities and disease associations. Methods: CRISPR sequences were retrieved from the CRISPR/Cas++ database, and human miRNA sequences were obtained from miRBase. Sequence alignments were performed using BLASTn with an E-value threshold of 1e-5 to identify significant similarities. Genes associated with matched human miRNAs were identified through the HGNC and GeneCards databases. Further analyses included comparison with disease-associated miRNAs reported in human and mouse datasets. Results: In Y. ruckeri, alignments revealed similarities to miRNAs linked with genes such as FOXO1, PTEN, PAX7, and DOCK3, which are associated with lung cancer and muscular dystrophies. In A. ambivalens, aligned miRNAs corresponded to loci including CHM13 and GRCh38, potentially linked to periembolic adenocarcinoma and mild pre-eclampsia. For Acidianus sp., matches were observed with miRNAs associated with genes like Irak2, NOS2, STAT1, and Numb, which have been implicated in Psoriatic arthritis, Alzheimer’s disease, Hepatocellular carcinoma, and Coronary artery di sease. Conclusion: CRISPR sequences from these prokaryotes show notable similarities with human miRNAs, suggesting possible indirect links to genes involved in major diseases. These preliminary findings emphasize the need for further investigation into shared sequence motifs and their functional roles in host-pathogen interactions or evolutionary biology.

Streptococci and Staphylococci are pathogenic agents that cause antibiotic-resistant infections through biofilm formation. Therefore, researchers are seeking alternative methods to combat antibiotic-resistant infections. This study aimed to compare the anti-biofilm effect of an aptamer-silver nanoparticle complex (Apt-AgNP) on Streptococci and Staphylococci. In the in silico studies, the physicochemical properties and secondary and tertiary structures of the selected bacterial surface proteins were compared and validated using ProtParam, GOR IV, SWISS-MODEL, Phyre2, I-TASSER, and GalaxyWEB servers. Aptamer binding to proteins was performed using molecular docking with HDock and ZDock servers. In the in vitro experiments, silver nanoparticles were synthesized and then attached to biotinylated AptBH via streptavidin. The anti-biofilm effect of Apt-AgNP on Streptococci and Staphylococci was compared with that of silver nanoparticles alone. For the characterization of silver nanoparticles and Apt-AgNP, XRD, FESEM, DLS, and zeta potential tests were used. The in silico results showed that aptamer docking with staphylococcal surface proteins yielded high binding scores, with the best results of − 310.74 for S. aureus and − 300.76 for S. epidermidis on the HDock server. Characterization results confirmed the spherical shape of the silver nanoparticles with a size of approximately 80 nm and their successful attachment to the aptamer. The Apt-AgNP at a concentration of 400 μg/mL showed a better anti-biofilm effect compared to silver nanoparticles alone. The highest anti-biofilm effect of this complex was observed on Staphylococci (69–72

Background: Food security has always been a concern in the multi-factorial systems analysis of health and wellbeing. The presence of nitrate and acrylamides in cooked meat leads to negative health outcomes. Objective: This study aimed to reduce nitrate and acrylamide content in different kinds of meats (chicken, turkey, lamb, beef, quail, and fish) using some plant extracts and lactobacillus treatment. Methods: The extracts were prepared from Coriandrum sativum, Nigella sativa, and Thymus leaves and stem. The used bacteria was Lactobacillus casei subsp. rhamnosus LCR6013. Acrylamide and nitrate were measured by liquid chromatography and colorimetric spectrophotometry methods Results: The results showed that both bacterial treatment and plant extracts could reduce the amount of acrylamide and nitrate. The most reduction in the amount of acrylamide and nitrate was obtained by adding Thymus and Nigella sativa extracts, followed by coriander extract and bacterial inoculum. Also, bacterial treatment was more effective for nitrate reduction than acrylamide. Conclusion: It was concluded that the plant extracts and bacterial treatment are appropriate solutions to reduce the amount of acrylamide and nitrate during the baking process of meat.

Haematococcus pluvialis is a type of microalgae that is commercially important as it is the primary source of natural astaxanthin - a potent antioxidant used in nutraceuticals, cosmetics, food, and aquaculture industries. Various nanoparticles and chemicals have been used to stimulate the growth of H. pluvialis to increase astaxanthin production. In this study, silicon nanoparticles were synthesized from tetraethyl orthosilicate (TEOS) and characterized to ensure the pure synthesis of uniform nanocrystals of silicon dioxide. The microalgae were cultivated in optimal Haematococcus medium (OHM) under specific conditions, including a temperature of 25 degrees C, a pH of 7, and a light intensity of 50 mu mol. s- 1.m- 2 for 12 h of light and 12 h of darkness. To study the amount of viability and astaxanthin production, the microalgae were exposed to different concentrations of silicon, sodium silicate, calcium silicate, and potassium silicate. The highest astaxanthin production (194 mg/g) was obtained after stimulation at 200 mu g/ml of silicon compared to the control after 15 days by high-performance liquid chromatography (HPLC) technique. Therefore, it can be concluded that certain concentrations of silicon and silicon salts can be considered a suitable stimulus to produce astaxanthin in the H. pluvialis microalgae. This study highlights the potential of using silicon nanoparticles as a stimulant for astaxanthin production in H. pluvialis, which could have significant implications for the nutraceuticals, cosmetics, food, and aquaculture industries.

Probiotics have poor gastrointestinal delivery because they lose their viability during intestinal passage. Microencapsulation has a significant effect on the survival of probiotic bacteria. This study aimed to synthesize chitosan-alginate nanoparticles and encapsulate Bacillus coagulans NBRC-12583 and Enterococcus faecium MGFR1 in alginate (Alg), chitosan (Cht) and inulin (Inu) by micro- and nano-encapsulation to investigate the viability of encapsulated strains in simulated gastrointestinal condition. The survival of free and encapsulated bacteria was studied for 120 min. The population of B. coagulans encapsulated in Cht-Alg only decreased by 0.86 log CFU, while that of E. faecium encapsulated in Cht-Alg nanoparticles only decreased by 0.27 log CFU. Treated with simulated intestinal fluid, B. coagulans and E. faecium populations decreased by 1.03 and 0.17 log CFU, respectively. E. faecium microencapsulated in Cht-Alg and Inu had a maximum encapsulation efficiency of 96.78%. E. faecium encapsulated in Cht-Alg nanoparticles and Inu was more viable than other encapsulated bacteria. Scanning electron microscopy was done to investigate the microcapsules' surface morphology, structure, internal cross-sectional view, and the zeta potential for the surface charge of Cht-Alg nanoparticles. The viability of the probiotic bacteria was enhanced significantly by microencapsulation and nanomaterial-based encapsulation within the chitosan-alginate and inulin matrix.

The development of a cancer vaccine has become an essential focus in the field of medical biotechnology and immunology. In our study, the NY-SAR-35 cancer/testis antigen was targeted to design a novel peptide vaccine using bioinformatics tools, and BALB/c mice were used to evaluate the vaccine’s immunological function. This evaluation involved assessing peptide-specific IgG levels in the serum via ELISA and measuring the levels of IFN-γ, IL-4, and granzyme B in the supernatant of cultured splenocytes. The final vaccine construct consisted of two T lymphocyte epitopes linked by the AAY linker. This construct displayed high antigenicity, non-allergenicity, non-toxicity, stability, and ability to induce IFN-γ and IL-4. It showed stable dynamics with both human MHC-I and II molecules, as well as mouse MHC-II molecules, and revealed strong Van der Waals and electrostatic energies. Emulsifying our peptide vaccine in incomplete Freund’s adjuvant resulted in a remarkable increase in the levels of IgG. The splenocytes of mice that received the combination of peptide and adjuvant displayed a noteworthy increase in IFN-γ, IL-4, and granzyme B secretion. Additionally, their lymphocytes exhibited higher proliferation rates compared to the control group. Our data demonstrated that our vaccine could stimulate a robust immune response, making it a promising candidate for cancer prevention. However, clinical trials are necessary to assess its efficacy in humans.

Bovine viral diarrhea virus (BVDV) is the cause of bovine viral diarrhea disease, one of the most economically important livestock diseases worldwide. The majority of BVD disease control programs rely on the detection and then elimination of persistent infection (PI) cattle, as the continuing source of disease. The main purpose of this study was to design and develop an accurate G-quadruplex-based aptasensor for rapid and simple detection of BVDV-1. In this work, we utilized in silico techniques to design a G-quadruplex aptamer specific for the detection of BVDV-1. Also, the rationally designed aptamer was validated experimentally and was used for developing a colorimetric biosensor based on an aptamer-gold nanoparticle system. Firstly, a pool of G-quadruplex forming ssDNA sequences was constructed. Then, based on the stability score in secondary and tertiary structures and molecular docking score, an aptamer (Apt31) was selected. In the experimental part, gold nanoparticles (AuNPs) with an average particle size of 31.7 nm were synthesized and electrostatically linked with the Apt31. The colorimetric test showed that salt-induced color change of AuNPs from red to purple-blue occurs only in the presence of BVDV-Apt31 complex, after 20 min. These results approved the specificity of Apt31 for BVDV. Furthermore, our biosensor could detect the virus at as low as 0.27 copies/ml, which is an acceptable value in comparison to the qPCR method. The specificity of the aptasensor was confirmed through cross-reactivity testing, while its selectivity was confirmed through plasma testing. The sample analysis showed 90% precision and 94% accuracy. It was concluded that the biosensor was adequately sensitive and specific for the detection of BVDV in plasma samples and could be used as a simple and rapid method on the farm.

Serine proteases are an essential and immensely diverse group of enzymes found in many different organisms, from mammals to viruses. Alkaline serine proteases (ASPs) are a type of serine protease that exhibit their highest activity levels in alkaline conditions. These enzymes play a critical role across a wide range of industries, providing immense value and benefits. ASPs are produced mainly by bacteria and fungi on industrial scales. The present study involved an analysis of various sequences of alkaline serine proteases derived from fungi and bacteria. The analytical approach employed encompassed the assessment of the pseudo amino acid composition (PseAAC), the tripeptide composition (TPC), physicochemical properties, secondary structures, and conserved motifs. Motif discovery and analysis showed that a considerable majority of bacterial alkaline serine protease sequences (over 94%) and fungal alkaline serine protease sequences (99%) in the dataset were associated with the subtilisin-like serine protease superfamily. This finding highlights the prevalence of this particular superfamily in alkaline serine protease sequences and provides valuable insight into the evolutionary relationships between different protease families. Based on the results of the study, the utilization of PseAAC and TPC techniques was successful in categorizing fungal and bacterial ASPs into separate groups. This was made possible by precise predictive models generated using machine learning algorithms. Bacterial and fungal ASPs had no significant differences in amino acid composition, ProtParam features, and GORIV secondary structure prediction outcomes. This underscores the importance of TPC and PseAAC concepts in accurately clustering and predicting ASP sequences.

More than 70% of hospital-acquired urinary tract infections are related to urinary catheters, which are commonly used for the treatment of about 20% of hospitalized patients. Urinary catheters are used to drain the bladder if there is an obstruction in the tube that carries urine out of the bladder (urethra). During catheter-associated urinary tract infections, microorganisms rise up in the urinary tract and reach the bladder, and cause infections. Various materials are used to fabricate urinary catheters such as silicone, polyurethane, and latex. These materials allow bacteria and fungi to develop colonies on their inner and outer surfaces, leading to bacteriuria or other infections. Urinary catheters could be modified to exert antibacterial and antifungal effects. Although so many research have been conducted over the past years on the fabrication of antibacterial and antifouling catheters, an ideal catheter needs to be developed for long-term catheterization of more than a month. In this review, we are going to introduce the recent advances in fabricating antibacterial materials to prevent catheter-associated urinary tract infections, such as nanoparticles, antibiotics, chemical compounds, antimicrobial peptides, bacteriophages, and plant extracts. image

L-asparaginase is a commercial enzyme with a wide variety of applications. Asparaginase is known as an anti-cancer agent that is effective for the treatment of certain lymphomas and leukemias by growth inhibition of human cancer cells. Additionally, asparaginase is used in the food industry in a pretreatment process to decrease the accumulation of carcinogenic acrylamide. In this paper, different aspects of bacterial and fungal asparaginases such as mass, hydrophobicity and hydrophilicity of pseudo amino acid composition (PseAAC), physicochemical properties, and structural motifs were studied, and ROC curve statistical analysis was used for the comparison. The results showed that none of the physicochemical properties of fungal and bacterial asparaginase could not be differed, except molecular weight and sequence length. MEME Suite analysis demonstrated that there was a motif that was specific for bacterial asparaginases. However, analysis based on the concept of PseACC indicated a differentiation line between fungal and bacterial asparaginases. In conclusion, although there was not any specific demonstration to separate the bacterial and fungal asparaginases in the case of physicochemical properties, PseAAC analysis can be an appropriate and usable method to differentiate between them.

One of the burning issues facing healthcare organizations is multidrug-resistant (MDR) bacteria. P. aeruginosa is an MDR opportunistic bacterium responsible for nosocomial and fatal infections in immunosuppressed individuals. According to previous studies, efflux pump activity and biofilm formation are the most common resistance mechanisms in P. aeruginosa. The aim of this study was to propose new antimicrobial peptides (AMPs) that target P. aeruginosa and can effectively address these resistance mechanisms through in silico and in vitro assessments. Since AMPs are an attractive alternative to antibiotics, in vitro experiments were carried out along with bioinformatics analyses on 19 Nef peptides (derived from the HIV1 Nef protein) in the current study. Several servers, including Dbaasps, Antibp2, CLASSAMP2, ToxinPred, dPABBs and ProtParam were used to predict Nef peptides as AMPs. To evaluate the binding affinities, a molecular docking analysis was performed with the HADDOCK web server for all Nef peptide models against two effective proteins of P. aeruginosa (MexB and PqsR) that play a role in efflux and quorum sensing. Moreover, the antibacterial and antibiofilm activity of the Nef peptides was investigated in a resistant strain of P. aeruginosa. The results of molecular docking revealed that all Nef peptides have a significant binding affinity to the abovementioned proteins. Nef-Peptide-19 has the highest affinity to the active sites of MexB and PqsR with the HADDOCK scores of -136.1 +/- 1.7 and -129.4 +/- 2, respectively. According to the results of in vitro evaluation, Nef peptide 19 showed remarked activity against P. aeruginosa with minimum inhibitory and bactericidal concentrations (MIC and MBC) of 10 mu M and 20 mu M, respectively. In addition, biofilm inhibitory activity was observed at a concentration of 20 mu M. Finally, Nef peptide 19 is proposed as a new AMP against P. aeruginosa.

Cancer is one of the main causes of death in the world. Resistance to anticancer treatments in patients with advanced solid tumors leads to new treatments. Therefore, more alternative anticancer methods have been found over time with greater specificity against tumor cells and with less or no adverse effects on normal cells. Bacterial spores of obligate anaerobes exclusively germinate in the hypoxic/necrotic areas and not in the well oxygenated areas of the body. This unique phenomenon has been exploited in using bacterial spores as a remedy for cancer. Bacterial toxins also play a significant role in either directly killing tumor cells or altering the cellular processes of the tumor cells which ultimately leads to the inhibition and regression of the solid tumor. In the microbial environment, pathogens such as Staphylococcus aureus, Bacillus cereus, or Streptococcus pyogenes produce hemolysin. This protein is used as an anti-cancer protein. To identify the production of hemolysin by bacteria, which can destroy cancer cells more effectively, different bacterial strains were first cultured in blood agar culture medium. The Strains that completely lysed red blood cells, creating transparent zones, were selected for further investigation. Then, to find out which strains have more ability to lyse red blood cells, the qualitative method of halo diameter measurement was used. Also, using quantitative methods, hemolysin strength in microtubes was determined compared to control samples. The results of the hemolysis in the microtube and the qualitative test results showed similar results. In the next step, the cell viability test was performed with the partially purified proteins. Then, bioinformatics studies such as secondary structure investigation, physicochemical properties, pseudo amino acid composition, and molecular docking were performed. The results of molecular docking showed that the hemolysin protein has the highest affinity for the cholesterol of the cytoplasmic membrane, respectively, of Bacillus subtilis, Bacillus cereus, and Staphylococcus aureus bacteria which play a significant role in either directly killing tumor cells or altering the cellular processes of the tumor cells which ultimately leads to the inhibition and regression of the solid tumor.